Heparin-aminobenzoate compositions



United States Patent HEP-AMINOBENZOATE COMPOSITIONS Marvin JayFahrenbach, Pearl River, and Kenneth Austen Burke, Nanuet, N. Y.,assignors to American Cg'ltc lnamid Company, New York, N. Y., acorporation 0 alne No Drawing. Application November 3, 1951, Serial No.254,809

4 Claims. (Cl. 167-74) This invention relates to new compositions ofmatter which have useful therapeutic efiects. The invention includes thenew substances, preparations containing the same, their use and methodsby which they are prepared.

In recent years certain sulfated polysaccharides, notably heparin, havehad an increased use in medicine in the treatment of diseases of thevascular system. These substances have been known for some time to beanti-coagulants and when injected into an animal increase the bloodclotting time. It has also been found that heparin and relatedanti-coagulants have an effect on the lipoproteins in the bloodstream;in pathological conditions low density lipo-proteins are thus convertedto higher density molecules, which reduces the tendency to form depositsof these materials in the vascular system. For this latter purpose, itis desirable that relatively low levels of the anti-coagulants bepresent in the bloodstream over a considerable period of time.

The anti-coagulant action of heparin and related anticoagulants is foundto be of comparatively short duration and in injection of 50 to 100 mg.of heparin, for example, in an adult patient may lose its effect onbloodclotting in 5 or 6 hours due to the fact that it is rapidlyexcreted or broken down in the body. Although some progress inprolonging the action of heparin following injection has been made byincorporating it in a menstruum which delays the absorption, yet it isfound that these are only partially successful and unduly painful to thepatient. As a result, many patients are reluctant to continue medicationwith the drug and the full advantages of such therapy cannot begenerally realized.

We have discovered that heparin and other sulfated polysaccharideshaving anti-coagulant activity can be reacted with certain localanesthetics of the amino-benzoate type to form insoluble compounds whichcan be injected in the patient with less pain. Of perhaps greaterimportance is thesurprising discovery that these compounds maintain atherapeutic level of the anti-coagulant in the bloodstream of sufficienteflect to allow the lipoproteins to reach their normal density, and thiseffect is maintained for a matter of days. Accordingly, it is possibleby the use of the compositions of the present invention to maintain inthe patient satisfactory levels of the drugs with a single relativelypainless injection the effects of which last for several days. Thisenables the clotting time of the blood to be prolonged for a protractedperiod and also enables the efiect of the heparin on the lipoproteins tobe maintained.

The anti-coagulant sulfated polysaccharides which may be reacted inaccordance with the present invention to provide new compositions ofmatter are generally of high molecular weight. Several of these arecommercially available and their use in the process of the presentinvention is shown in specific examples which follow.

The amino benzoates which may be reacted with the sulfatedanti-coagulants have the following general formula:

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To a solution of 1 part sodium hepalinate in 25 volumes of water at 25C. was slowly added a solution of 3 parts S-dibutylaminopropylp-aminobenzoate sulfate, sold un der the name Butacaine sulfate, involumes of water. A white, finely divided precipitate formedimmediately. The pH of this slurry was about 5.0. After stirring for 10minutes at 25 C., the white solid was filtered on a sintered glassfunnel, washed with 50 volumes of water, them redissolved in 200 volumesof 50% alcohol. This solution had a colloidal appearance. Afterfiltration of the solution, the filtrate and wash (250 volumes) stillremained slightly colloidal in appearance. By drying an aliquot, it wasshown that there were 3 parts of the Butacaine-heparin salt present.

Dilution of one part of the above solution to 18 and 180 parts by volumewith water resalted in a solution which contained 0.00067 and 0.000067part respectively of heparin-butacaine per volume. Injection of 0.5volume of these two solutions into white rats once a day for 3 daysresulted in no gross toxicity. By the virtue of the high molecularweight and variable composition of heparin, the products that areobtained in accordance with the above procedure and in other examplesare not sharply characteristic in physical properties such as meltingpoint or crystalline structure.

Example 2 To a solution of 5 parts sodium heparinate in 200 volumes ofwater at 45 C. was slowly added a solution of 15 parts Butacaine sulfatein 400 volumes of water; a cream colored finely divided solidprecipitated immediately. This solid was separated from the supernatantby centrifugation for 15 minutes at 1850 R. P. M. The deep cream coloredresidue was washed three times with 150 volumes of water, each timeremoving the supernatant by centrifugation for 15 minutes at 1850 R. P.M. Weight of the wet residue of Butacaine-heparin was 30.6 parts. A 5.2part aliquot of this residue was dried in a vacuum desiccator overconcentrated sulfuric acid to give 1.6 parts of a light tan coloredsolid which pulverized easily to a fine particle size. The productmelted to a viscous oil between -125 C.

Example 3 To a solution of 1 part of the sulfuric acid ester ofpolyanhydromannuronic acid, sold under the name Paritol- A, in 25volumes of water at 25 C. was slowly added with stirring a solution of 3parts Butacaine sulfate in 25 volumes of water. A heavy white finelydivided precipitate formed immediately and settled quickly. This whiteButacaine-Paritol-A salt was filtered on a sintered glass funnel, washedwith 50 volumes of water, and dried in a vacuum desiccator overanhydrous magnesium sulfate to give 2.61 parts of a hard vitreous solid,which was readily pulverized in a mortar and pestle to a fine whitepowder. The product melted to a viscous oil between 124-140" C. withdecomposition.

. Example 4 To 0.05 part of the sodium salt of sulfated polygalacturonicacid methyl ester methyl glycoside, sold under the name Treburon, in 2volumes of 0.9 sodium chloride solution was slowly added 3.08 volumes of0.1 N silver nitrate solution in order to remove chloride ions. Theprecipitate of silver chloride was removed by centrifugation. To thesupernatant solution of-Treburon at 25 C. was slowly added 0.15 part ofButacaine sulfate in 5 volumes -of Water. A white, finely divided solidprecipitated. This Butacaine-Treburon salt was filtered on a sinteredglass funnel, washed with "20 volumes of water and dried in a vacuumdesiccator over anhydrous magnesium sulfate to give 0.110 part of :ahard vitreous solid, which was readily pulverized in a mortar and pestleto a fine 'w'hite powder. The product melted to a viscous oi l between155-165 C. with slight decomposition.

Example 5 To a 2 volume portion of a 30% aqueous solution of sodiumheparinate was slowly added 2 volumes of a 30% aqueous solution of2-.dimethylaminoethyl p-butylaminobenzoate hydrochloride. .An oilycollodial solution was formed which changed slowly to a gummy veryfinely divided solid after standing at room temperature. The solids wereseparated by centrifugation, washed twice with volumes of water anddried in :a vacuum desiccator over anhydrous magnesium sulfate to agummy vitreous solid.

Example 6 To .a 2 volume portion of va 30% aqueous solution .of sodiumheparinate is slowly :added 2 volumes of a 30% aqueous solution of.S-dimethylamino-LZ-dipropyl paminobenzoate hydrochloride. .An oilycollodial :solution was formed which .changed to a gummy very timelydivided solid aliter standing .at room temperature. The solid wasseparated by centrifugation, washed twice with '10 volumes of water anddried .in a vacuum desiccator over anhydrous magnesium sulfate "to .agummy vitreous solid.

What we claimis:

1. A new composition of matter comprising the solid product obtained byreacting substantially equivalent amounts of heparin and anaminobenzoate local anesthetic having the following formula ReferencesCited in the file of this patent UNITED STATES PATENTS 2,100,054.Hopkinson Nov. 23, 1937 2,487,975 .Koree Nov. 15, 1949 2,508,433 SnyderMay 23, 1950 2,561,384 Lee July 24, 1951 FOREIGN PATENTS 653,368 GreatBritain May 16, 1951 OTHER REFERENCES Loomis: I. Pharmacol. and Exptl.Therap., vol. 104,

No. 1, pp. 87-92, January 1952. (Recdfor publication Oct. 8., 1951.)(Copy in POSL 167-65.5A').

Vorzimer: J. A. M. A., vol. 138, No. 10, pp. 747-8. Clinical Medicine,August 1948, p. 197.

'Punde'l: Presse Med, vol. 57 ('Dec. 3, 1949), pp. 1120-1.

I. A. M. A., vol. 139., No. 13, p. 883, March 26, 1949.

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.Proc. Stall Meetings Mayo Clinic, Dec. 10, 1947, pp. 567-570.

1. A NEW COMPOSITION OF MATTER COMPRISING THE SOLID PRODUCT OBTAINED BYREACTING SUBSTANTIALLY EQUIVALENT AMOUNTS OF HEPARIN AND ANAMINOBENZOATE LOCAL ANESTHETIC HAVING THE FOLLOWING FORMULA